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Method for producing marek disease live vaccine of chicken by using cell line

A technology of chicken Marek's disease and cell lines, which is applied in the field of production of chicken Marek's disease live vaccines with cell lines, can solve problems affecting vaccine production and quality, contamination of cells with exogenous viruses, and low vaccine potency, and achieve good economy Benefits and application prospects, high virus content, good immune efficacy

Active Publication Date: 2015-06-17
南京创启生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although my country has stipulated that chicken embryos and cells for preparing poultry vaccines must come from specific pathogen-free (SPF) chicken flocks, in reality, due to the lag of detection and the complexity of the poultry breeding environment in my country, chicken embryos for poultry vaccine production Cells are easy to cause cell-exogenous virus contamination, and the vaccine potency is not high, and the difference between batches is large, which seriously affects the yield and quality of the vaccine

Method used

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  • Method for producing marek disease live vaccine of chicken by using cell line
  • Method for producing marek disease live vaccine of chicken by using cell line
  • Method for producing marek disease live vaccine of chicken by using cell line

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Preparation of virus seeds: Take out the preserved CVI988 / Rispens virus seeds from liquid nitrogen, melt them quickly (within 1 minute) in warm water at 37°C, immediately dilute them in DF-1 cell growth medium at 1:5, and centrifuge at 1000r / min After 10 minutes, discard the supernatant, dilute with cell growth medium to contain 200,000 PFU / ml, inoculate 100ml square flask with DF-1 single layer, inoculate 0.5ml in each bottle, absorb at 36.5-37.5°C for 1 hour, add maintenance solution , cultured at 36.5-37.5°C in 5% CO2 for 2-4 days, when 70% of the monolayer showed cytopathic changes, and a large number of confluent cells and refraction round cells were produced, add 0.25% trypsin-0.02%EDTA (1:4) Digest and disperse the cells with the cell dispersion solution. After the harvested cell suspension is centrifuged at 1000r / min for 10 minutes, the supernatant is discarded, and the precipitated cells are suspended and inoculated with DF-1 monolayer with an appropriate am...

Embodiment 2

[0042] Propagation of virus seeds: the chicken Marek's disease type III turkey herpes virus Fc-126 basic seeds are diluted with seed virus (equal mixture of 2 times 199 and 2 times hydrolyzed milk protein solution of 5% newborn bovine serum, containing 100 Unit / ml double antibody, pH adjusted to 7.2) diluted to 400,000 PFU / ml, inoculated on well-grown DF-1 cells in a 0.5ml / 100ml square bottle, adsorbed at 36.5-37.5°C for 1 hour, and then added an appropriate amount of maintenance solution , continue to cultivate, when 70% and above monolayer cells have typical Marek's disease cell lesions, digest and disperse the cells with EDTA-trypsin cell dispersion solution, and the harvested cell suspension is used as a virus seed for seedling production;

[0043] Passage and culture of cells for seedling production: DF-1 cell line is digested and passaged with EDTA-trypsin cell dispersion liquid, and then cultured with cell growth medium at 36.5-37.5°C. When a good monolayer is formed, it...

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Abstract

The invention discloses a method for producing a marek disease live vaccine of chicken by using a cell line. The method comprises the following steps: A, selecting the cell line as a vaccine-preparing cell; B, passaging and cultivating the vaccine-preparing cell; C, breeding cytotoxic species; D, breeding vaccine-preparing venom; E, preparing vaccine, carrying out split charging, freeze-drying or cryopreserving in liquid nitrogen. By adopting the method, the defects of the prior art can be solved, the method has the advantages of being stable in production process, easy to operate, high in virus content, small in difference between batches, and easy in quality control, and the yield and quality of the vaccine can be significantly improved. The marek disease live vaccine of chicken produced by the method is good in safety, and high in immunity, and has complete immune protection action on attack of a virulent strain or a very virulent strain of chicken marek's disease.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for producing chicken Marek's disease live vaccine with cell lines. Background technique [0002] The cells used in the production of chicken Marek's disease live vaccines in my country are primary chicken embryo fibroblasts. Although my country has stipulated that chicken embryos and cells for preparing poultry vaccines must come from specific pathogen-free (SPF) chicken flocks, in reality, due to the lag of detection and the complexity of the poultry breeding environment in my country, chicken embryos for poultry vaccine production Cells are easy to cause cell-exogenous virus contamination, and the vaccine potency is not high, and the batch-to-batch variation is large, which seriously affects the yield and quality of the vaccine. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a metho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/245A61K39/255A61P31/22A61P35/00
Inventor 陆秀云
Owner 南京创启生物科技有限公司
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